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COMBINED METHODS TO EVALUATE HUMAN CELLS IN MUSCLE XENOGRAFTS
Author
Affilliation
Abstract
Xenotransplantation of human cells into immunodeficient mouse models is a very powerful
tool and an essential step for the pre-clinical evaluation of therapeutic cell- and gene- based
strategies. Here we describe an optimized protocol combining immunofluorescence and
real-time quantitative PCR to both quantify and visualize the fate and localization of human
myogenic cells after injection in regenerating muscles of immunodeficient mice. Whereas
real-time quantitative PCR-based method provides an accurate quantification of human
cells, it does not document their specific localization. The addition of an immunofluorescence
approach using human-specific antibodies recognizing engrafted human cells gives
information on the localization of the human cells within the host muscle fibres, in the stem
cell niche or in the interstitial space. These two combined approaches offer an accurate
evaluation of human engraftment including cell number and localization and should provide
a gold standard to compare results obtained either using different types of human stem cells
or comparing healthy and pathological muscle stem cells between different research laboratories
worldwide.
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